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Single-Molecule Imaging of Lateral Mobility and Ion Channel Activity in Lipid Bilayers using Total Internal Reflection Fluorescence (TIRF) Microscopy
JoVE 신문
생화학
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JoVE 신문 생화학
Single-Molecule Imaging of Lateral Mobility and Ion Channel Activity in Lipid Bilayers using Total Internal Reflection Fluorescence (TIRF) Microscopy
DOI:

08:55 min

February 17, 2023

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Chapters

  • 00:05Introduction
  • 00:43Preparation of Lipids and Agarose Hydrogel
  • 01:33PMMA Chamber Assembly and Lipid Monolayer Formation
  • 03:06Preparation of Lipid-Coated Aqueous Droplets in Hexadecane/Silicone Oil Solution
  • 04:17Preparation and Imaging of Single Ion Channels in DIB Membranes
  • 06:49Results: Isolation of TOM-CC and Correlation of Channel Activity with Lateral Mobility of TOM-CC
  • 08:18Conclusion

Summary

자동 번역

This protocol describes how to use TIRF microscopy to track individual ion channels and determine their activity in supported lipid membranes, thereby defining the interplay between lateral membrane movement and channel function. It describes how to prepare the membranes, record the data, and analyze the results.

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